MicroPure II Column
MicroPure II Column
Key featuresShow Hide
- High performance hydrophobic polymeric resin.
- Removes the most common impurities and truncated sequences
- The method is rapid, efficient and many samples can be purified simultaneously.
- At least 1 µmol or 50 O.D's of crude DNA can be applied to the column.
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MicroPure II columns are intended for reverse-phase purification of trityl-protected oligonucleotides followed by on-column detritylation. The cartridge consists of a 5 mL syringe barrel packed with 200 mg of high performance hydrophobic polymeric resin. The method relies on strong binding between the trityl-protected product and the resin, thus allowing separation from the most common impurities, truncated sequences, which do not bind. Treatment with acid removes the DMT group from the product, the desired product (free from organic residues) is eluted with 20% acetonitrile. The method is rapid, efficient and many samples can be purified simultaneously. At least 1 µmol or 50 O.D's of crude DNA can be applied to the column. The purified yield will depend, in each case, on sequence length and the quality of the crude sample.
- Program DNA synthesizer to leave terminal 5'-DMT group attached;
- Cleave and deprotect product with conc. aqueous ammonia;
- Evaporate cleavage solution to dryness;
- Re-suspend in 1ml 0.1N TEAA or H2O;
- Pre-equilibrate the column by passing 4mL of acetonitrile through the column;
- Repeat with 6mL of 1N TEAA (triethylamine acetate);
- Take the oligo solution of step 4 and apply slowly to the column while collecting effluent in a separate micro-centrifuge tube. Reload effluent and pass once more through column (optional);
- To elute failures pass 20 mL (2mL at a time) of 12% MeCN/ 0.1N TEAA through the column;
- Rinse the column with 6mL of water;
- Pass 4mL (2 at a time) of 2.5% TFA through the column, wait 5 minutes;
- Pass 6mL or more of water through column. Check to see pH is neutral.
- Slowly pass 10mL of 20% acetonitrile/water through the column, collecting effluent containing the desired purified product in a labeled microcentrifuge tube;
- Speedvac ™ the solution to dryness, analyze and use as desired.
Storage and handling:
- Shipping conditions: Ambient
- Storage conditions: Room Temperature